Secretory Tissues and Volatile Components of Disc Florets in Several Wild Helianthus L. Species.

Although flower pollinator interactions are known to be mediated by floral traits, not enough attention has been paid to the research of secretory tissues and volatile components of sunflower disc florets as potentially important parameters in breeding programs. (1) To our knowledge, this is the first integrated study aimed at better understanding the attractiveness of sunflower capitula to insects. In the study, we have made a very detailed comparative analysis of secretory tissues and the characterization of the volatile components (VOCs) of disc florets in 10 wild perennial Helianthus species. (2) For anatomical analyses, cross-sections were obtained from the nectary zone of disc florets using a cryotechnique procedure. Micromorphological observation and morphological and anatomical analysis of disc florets were performed using light and scanning electron microscopy. For VOCs, we applied headspace, GC-FID, and GC/MS analyses. (3) The obtained results indicate that there is a difference between the analyzed traits among studied species. H. eggertii, H. hirsutus, H. mollis, H. resinosus, and H. tuberosus had high disc diameter values, a high cross-section area and disc floret corolla length, as well as the largest cross-section area and thickness of the disc florets nectary. In the analyzed VOCs, 30 different compounds were detected. The highest yield and quantity of α-Pinene was observed in H. mollis. (4) Inflorescence features, such as receptacle diameter, corolla and secretory tissue properties, and floret VOCs production and characterization, provided valuable information that can be used as guidelines in sunflower breeding programs to maximize pollinator attractiveness and increase seed yield.

Genome editing for healthy crops: traits, tools and impacts.

Crop cultivars in commercial use have often been selected because they show high levels of resistance to pathogens. However, widespread cultivation of these crops for many years in the environments favorable to a pathogen requires durable forms of resistance to maintain "healthy crops". Breeding of new varieties tolerant/resistant to biotic stresses by incorporating genetic components related to durable resistance, developing new breeding methods and new active molecules, and improving the Integrated Pest Management strategies have been of great value, but their effectiveness is being challenged by the newly emerging diseases and the rapid change of pathogens due to climatic changes. Genome editing has provided new tools and methods to characterize defense-related genes in crops and improve crop resilience to disease pathogens providing improved food security and future sustainable agricultural systems. In this review, we discuss the principal traits, tools and impacts of utilizing genome editing techniques for achieving of durable resilience and a "healthy plants" concept.

Oil yield prediction for sunflower hybrid selection using different machine learning algorithms.

Due to the increased demand for sunflower production, its breeding assignment is the intensification of the development of highly productive oil seed hybrids to satisfy the edible oil industry. Sunflower Oil Yield Prediction (SOYP) can help breeders to identify desirable new hybrids with high oil yield and their characteristics using machine learning (ML) algorithms. In this study, we developed ML models to predict oil yield using two sets of features. Moreover, we evaluated the most relevant features for accurate SOYP. ML algorithms that were used and compared were Artificial Neural Network (ANN), Support Vector Regression, K-Nearest Neighbour, and Random Forest Regressor (RFR). The dataset consisted of samples for 1250 hybrids of which 70% were randomly selected and were used to train the model and 30% were used to test the model and assess its performance. Employing MAE, MSE, RMSE and R2 evaluation metrics, RFR consistently outperformed in all datasets, achieving a peak of 0.92 for R2 in 2019. In contrast, ANN recorded the lowest MAE, reaching 65 in 2018 The paper revealed that in addition to seed yield, the following characteristics of hybrids were important for SOYP: resistance to broomrape (Or) and downy mildew (Pl) and maturity. It was also disclosed that the locality feature could be used for the estimation of sunflower oil yield but it is highly dependable on weather conditions that affect the oil content and seed yield. Up to our knowledge, this is the first study in which ML was used for sunflower oil yield prediction. The obtained results indicate that ML has great potential for application in oil yield prediction, but also selection of parental lines for hybrid production, RFR algorithm was found to be the most effective and along with locality feature is going to be further evaluated as an alternative method for genotypic selection.

Exploitation of epigenetic variation of crop wild relatives for crop improvement and agrobiodiversity preservation.

Crop wild relatives (CWRs) are recognized as the best potential source of traits for crop improvement. However, successful crop improvement using CWR relies on identifying variation in genes controlling desired traits in plant germplasms and subsequently incorporating them into cultivars. Epigenetic diversity may provide an additional layer of variation within CWR and can contribute novel epialleles for key traits for crop improvement. There is emerging evidence that epigenetic variants of functional and/or agronomic importance exist in CWR gene pools. This provides a rationale for the conservation of epigenotypes of interest, thus contributing to agrobiodiversity preservation through conservation and (epi)genetic monitoring. Concepts and techniques of classical and modern breeding should consider integrating recent progress in epigenetics, initially by identifying their association with phenotypic variations and then by assessing their heritability and stability in subsequent generations. New tools available for epigenomic analysis offer the opportunity to capture epigenetic variation and integrate it into advanced (epi)breeding programmes. Advances in -omics have provided new insights into the sources and inheritance of epigenetic variation and enabled the efficient introduction of epi-traits from CWR into crops using epigenetic molecular markers, such as epiQTLs.

Crop breeding for a changing climate in the Pannonian region: towards integration of modern phenotyping tools.

The Pannonian Plain, as the most productive region of Southeast Europe, has a long tradition of agronomic production as well as agronomic research and plant breeding. Many research institutions from the agri-food sector of this region have a significant impact on agriculture. Their well-developed and fruitful breeding programmes resulted in productive crop varieties highly adapted to the specific regional environmental conditions. Rapid climatic changes that occurred during the last decades led to even more investigations of complex interactions between plants and their environments and the creation of climate-smart and resilient crops. Plant phenotyping is an essential part of botanical, biological, agronomic, physiological, biochemical, genetic, and other omics approaches. Phenotyping tools and applied methods differ among these disciplines, but all of them are used to evaluate and measure complex traits related to growth, yield, quality, and adaptation to different environmental stresses (biotic and abiotic). During almost a century-long period of plant breeding in the Pannonian region, plant phenotyping methods have changed, from simple measurements in the field to modern plant phenotyping and high-throughput non-invasive and digital technologies. In this review, we present a short historical background and the most recent developments in the field of plant phenotyping, as well as the results accomplished so far in Croatia, Hungary, and Serbia. Current status and perspectives for further simultaneous regional development and modernization of plant phenotyping are also discussed.

KASP Markers Specific for the Fertility Restorer Locus Rf1 and Application for Genetic Purity Testing in Sunflowers (Helianthus annuus L.).

Single nucleotide polymorphisms (SNPs) were significantly associated with fertility restoration of cytoplasmic male sterility (CMS) PET1 by the restorer gene Rf1. For these SNPs, four Kompetitive allele-specific PCR (KASP) markers were successfully designed. The KASP markers cover the fertility restorer locus Rf1, spanning about 3 Mb, and clearly differentiate restorer and maintainer lines. For genetic purity testing in sunflower hybrid production, the efficiency for detecting contaminations in samples was simulated using mixtures of hypocotyls or leaves. Contaminations of restorer lines with 1%, 3%, 5%, 10%, and 50% of maintainer lines were screened with all four KASP markers. Contaminations of 10% could be clearly detected in pools of 100 plants. Contaminations below this level require detection on a single plant level. For single plant detections, ethyl methanesulfonate-treated sunflower F1 hybrids, which had been phenotypically evaluated for male sterility (potential mutation in the Rf1 gene) were screened. Nine identified either partially male-sterile or male-sterile plants were analyzed with all four KASP markers and only one proved to be a hybrid with a mutation, seven were male-sterile contaminants in the F1 seeds used (1.6%) and one a recombinant plant. The four KASP markers should be valuable tools for marker-assisted selection (MAS) in sunflower breeding regarding the restorer locus Rf1.

An Epigenetic Alphabet of Crop Adaptation to Climate Change.

Crop adaptation to climate change is in a part attributed to epigenetic mechanisms which are related to response to abiotic and biotic stresses. Although recent studies increased our knowledge on the nature of these mechanisms, epigenetics remains under-investigated and still poorly understood in many, especially non-model, plants, Epigenetic modifications are traditionally divided into two main groups, DNA methylation and histone modifications that lead to chromatin remodeling and the regulation of genome functioning. In this review, we outline the most recent and interesting findings on crop epigenetic responses to the environmental cues that are most relevant to climate change. In addition, we discuss a speculative point of view, in which we try to decipher the "epigenetic alphabet" that underlies crop adaptation mechanisms to climate change. The understanding of these mechanisms will pave the way to new strategies to design and implement the next generation of cultivars with a broad range of tolerance/resistance to stresses as well as balanced agronomic traits, with a limited loss of (epi)genetic variability.

Genetic Improvement in Sunflower Breeding-Integrated Omics Approach.

Foresight in climate change and the challenges ahead requires a systematic approach to sunflower breeding that will encompass all available technologies. There is a great scarcity of desirable genetic variation, which is in fact undiscovered because it has not been sufficiently researched as detection and designing favorable genetic variation largely depends on thorough genome sequencing through broad and deep resequencing. Basic exploration of genomes is insufficient to find insight about important physiological and molecular mechanisms unique to crops. That is why integrating information from genomics, epigenomics, transcriptomics, proteomics, metabolomics and phenomics enables a comprehensive understanding of the molecular mechanisms in the background of architecture of many important quantitative traits. Omics technologies offer novel possibilities for deciphering the complex pathways and molecular profiling through the level of systems biology and can provide important answers that can be utilized for more efficient breeding of sunflower. In this review, we present omics profiling approaches in order to address their possibilities and usefulness as a potential breeding tools in sunflower genetic improvement.

Deciphering the Epigenetic Alphabet Involved in Transgenerational Stress Memory in Crops.

Although epigenetic modifications have been intensely investigated over the last decade due to their role in crop adaptation to rapid climate change, it is unclear which epigenetic changes are heritable and therefore transmitted to their progeny. The identification of epigenetic marks that are transmitted to the next generations is of primary importance for their use in breeding and for the development of new cultivars with a broad-spectrum of tolerance/resistance to abiotic and biotic stresses. In this review, we discuss general aspects of plant responses to environmental stresses and provide an overview of recent findings on the role of transgenerational epigenetic modifications in crops. In addition, we take the opportunity to describe the aims of EPI-CATCH, an international COST action consortium composed by researchers from 28 countries. The aim of this COST action launched in 2020 is: (1) to define standardized pipelines and methods used in the study of epigenetic mechanisms in plants, (2) update, share, and exchange findings in epigenetic responses to environmental stresses in plants, (3) develop new concepts and frontiers in plant epigenetics and epigenomics, (4) enhance dissemination, communication, and transfer of knowledge in plant epigenetics and epigenomics.

Targeted plant improvement through genome editing: from laboratory to field.

KEY MESSAGE: This review illustrates how far we have come since the emergence of GE technologies and how they could be applied to obtain superior and sustainable crop production. The main challenges of today's agriculture are maintaining and raising productivity, reducing its negative impact on the environment, and adapting to climate change. Efficient plant breeding can generate elite varieties that will rapidly replace obsolete ones and address ongoing challenges in an efficient and sustainable manner. Site-specific genome editing in plants is a rapidly evolving field with tangible results. The technology is equipped with a powerful toolbox of molecular scissors to cut DNA at a pre-determined site with different efficiencies for designing an approach that best suits the objectives of each plant breeding strategy. Genome editing (GE) not only revolutionizes plant biology, but provides the means to solve challenges related to plant architecture, food security, nutrient content, adaptation to the environment, resistance to diseases and production of plant-based materials. This review illustrates how far we have come since the emergence of these technologies and how these technologies could be applied to obtain superior, safe and sustainable crop production. Synergies of genome editing with other technological platforms that are gaining significance in plants lead to an exciting new, post-genomic era for plant research and production. In previous months, we have seen what global changes might arise from one new virus, reminding us of what drastic effects such events could have on food production. This demonstrates how important science, technology, and tools are to meet the current time and the future. Plant GE can make a real difference to future sustainable food production to the benefit of both mankind and our environment.

Digital Image Analysis Using FloCIA Software for Ornamental Sunflower Ray Floret Color Evaluation.

As an esthetic trait, ray floret color has a high importance in the development of new sunflower genotypes and their market value. Standard methodology for the evaluation of sunflower ray florets is based on International Union for the Protection of New Varieties of Plants (UPOV) guidelines for sunflower. The major deficiency of this methodology is the necessity of high expertise from evaluators and its high subjectivity. To test the hypothesis that humans cannot distinguish colors equally, six commercial sunflower genotypes were evaluated by 100 agriculture experts, using UPOV guidelines. Moreover, the paper proposes a new methodology for sunflower ray floret color classification - digital UPOV (dUPOV), that relies on software image analysis but still leaves the final decision to the evaluator. For this purpose, we created a new Flower Color Image Analysis (FloCIA) software for sunflower ray floret digital image segmentation and automatic classification into one of the categories given by the UPOV guidelines. To assess the benefits and relevance of this method, accuracy of the newly developed software was studied by comparing 153 digital photographs of F2 genotypes with expert evaluator answers which were used as the ground truth. The FloCIA enabled visualizations of segmentation of ray floret images of sunflower genotypes used in the study, as well as two dominant color clusters, percentages of pixels belonging to each UPOV color category with graphical representation in the CIE (International Commission on Illumination) L∗a∗b∗ (or simply Lab) color space in relation to the mean vectors of the UPOV category. Precision (repeatability) of ray flower color determination was greater between dUPOV based expert color evaluation and software evaluation than between two UPOV based evaluations performed by the same expert. The accuracy of FloCIA software used for unsupervised (automatic) classification was 91.50% on the image dataset containing 153 photographs of F2 genotypes. In this case, the software and the experts had classified 140 out of 153 of images in the same color categories. This visual presentation can serve as a guideline for evaluators to determine the dominant color and to conclude if more than one significant color exists in the examined genotype.

Epigenetics: possible applications in climate-smart crop breeding.

To better adapt transiently or lastingly to stimuli from the surrounding environment, the chromatin states in plant cells vary to allow the cells to fine-tune their transcriptional profiles. Modifications of chromatin states involve a wide range of post-transcriptional histone modifications, histone variants, DNA methylation, and activity of non-coding RNAs, which can epigenetically determine specific transcriptional outputs. Recent advances in the area of '-omics' of major crops have facilitated identification of epigenetic marks and their effect on plant response to environmental stresses. As most epigenetic mechanisms are known from studies in model plants, we summarize in this review recent epigenetic studies that may be important for improvement of crop adaptation and resilience to environmental changes, ultimately leading to the generation of stable climate-smart crops. This has paved the way for exploitation of epigenetic variation in crop breeding.

Genetic and Genomic Tools in Sunflower Breeding for Broomrape Resistance.

Broomrape is a root parasitic plant causing yield losses in sunflower production. Since sunflower is an important oil crop, the development of broomrape-resistant hybrids is the prime breeding objective. Using conventional plant breeding methods, breeders have identified resistant genes and developed a number of hybrids resistant to broomrape, adapted to different growing regions worldwide. However, the spread of broomrape into new countries and the development of new and more virulent races have been noted intensively. Recent advances in sunflower genomics provide additional tools for plant breeders to improve resistance and find durable solutions for broomrape spread and virulence. This review describes the structure and distribution of new, virulent physiological broomrape races, sources of resistance for introduction into susceptible cultivated sunflower, qualitative and quantitative resistance genes along with gene pyramiding and marker assisted selection (MAS) strategies applied in the process of increasing sunflower resistance. In addition, it presents an overview of underutilized biotechnological tools, such as phenotyping, -omics, and genome editing techniques, which need to be introduced in the study of sunflower resistance to broomrape in order to achieve durable resistance.

Author Correction: Increase of isoflavones in the aglycone form in soybeans by targeted crossings of cultivated breeding material.

An amendment to this paper has been published and can be accessed via a link at the top of the paper.

Increase of isoflavones in the aglycone form in soybeans by targeted crossings of cultivated breeding material.

Isoflavones are a group of phytoestrogens, naturally-occurring substances important for their role in human health. Legumes, particularly soybeans (Glycine max (L.) Merr.), are the richest source of isoflavones in human diet. Since there is not much current data on genetics of isoflavones in soybean, particularly in the aglycone form, elucidation of the mode of inheritance is necessary in order to design an efficient breeding strategy for the development of high-isoflavone soybean genotypes. Based on the isoflavone content in 23 samples of soybeans from four different maturity groups (00, 0, I and II), three crosses were made in order to determine the inheritance pattern and increase the content of total isoflavones and their aglycone form. Genotype with the lowest total isoflavone content (NS-L-146) was crossed with the low- (NS Zenit), medium (NS Maximus), and high- (NS Virtus) isoflavone genotypes. There were no significant differences in the total isoflavone content (TIF) between F2 populations, and there was no transgression among genotypes within the populations. Each genotype within all three populations had a higher TIF value than the lower parent (NS-L-146), while genotypes with a higher TIF value than the better parent were found only in the NS-L-146 × NS Zenit cross. However, significant differences in the aglycone ratio (ratio of aglycone to glycone form of isoflavones) were found between the populations. The highest aglycone ratio was found in the NS-L-146 × NS Maximus cross. The results indicate that the genetic improvement for the trait is possible.

Development and Validation of Markers for the Fertility Restorer Gene Rf1 in Sunflower.

Hybrid breeding in sunflowers based on CMS PET1 requires development of restorer lines carrying, in most cases, the restorer gene Rf1. Markers for marker-assisted selection have been developed, but there is still need for closer, more versatile, and co-dominant markers linked to Rf1. Homology searches against the reference sunflower genome using sequences of cloned markers, as well as Bacterial Artificial Chromosome (BAC)-end sequences of clones hybridizing to them, allowed the identification of two genomic regions of 30 and 3.9 Mb, respectively, as possible physical locations of the restorer gene Rf1 on linkage group 13. Nine potential candidate genes, encoding six pentatricopeptide repeat proteins, one tetratricopeptide-like helical domain, a probable aldehyde dehydrogenase 22A1, and a probable poly(A) polymerase 3 (PAPS3), were identified in these two genomic regions. Amplicon targeted next generation sequencing of these nine candidate genes for Rf1 was performed in an association panel consisting of 27 maintainer and 32 restorer lines and revealed the presence of 210 Single Nucleotide Polymorphisms (SNPs) and 67 Insertions/Deletions (INDELs). Association studies showed significant associations of 10 SNPs with fertility restoration (p-value < 10-4), narrowing Rf1 down to three candidate genes. Three new markers, one co-dominant marker 67N04_P and two dominant markers, PPR621.5R for restorer, and PPR621.5M for maintainer lines were developed and verified in the association panel of 59 sunflower lines. The versatility of the three newly developed markers, as well as of three existing markers for the restorer gene Rf1 (HRG01 and HRG02, Cleaved Amplified Polymorphic Sequence (CAPS)-marker H13), was analyzed in a large association panel consisting of 557 accessions.

Sunflower Genetics from Ancestors to Modern Hybrids-A Review.

Domestication and the first steps of sunflower breeding date back more than 4000 years. As an interesting crop to humans, sunflower underwent significant changes in the past to finally find its place as one of the most significant oil crops today. Substantial progress has already been made in understanding how sunflower was domesticated. Recent advances in molecular techniques with improved experimental designs contributed to further understanding of the genetic and molecular basis underlying the architectural and phenotypic changes that occurred during domestication and improvements in sunflower breeding. Understanding the domestication process and assessing the current situation concerning available genotypic variations are essential in order for breeders to face future challenges. A review of the tools that are used for exploring the genetic and genome changes associated with sunflower domestication is given in the paper, along with a discussion of their possible implications on classical sunflower breeding techniques and goals.